[Efficiency of CNV-seq in detecting fetal DMD gene deletion or duplication in prenatal diagnosis].

Objective: To evaluate the diagnostic efficiency of copy number variation sequencing (CNV-seq) to detect the deletion or duplication of DMD gene in prenatal diagnosis. Methods: A retrospective analysis was carried out on the CNV-seq results of 34 544 fetuses diagnosed in the First People's Hospital of Yunnan Province from January 2018 to July 2023. A total of 156 cases of fetuses were collected, including Group 1:125 cases with family history of Duchenne muscular dystrophy or Becker muscular dystrophy (DMD/BMD), and Group 2:31 cases with no family history but a DMD gene deletion or duplication was detected unexpectedly by CNV-seq. Multiplex ligation-dependent probe amplification (MLPA) was used as a standard method to detect the deletion or duplication. Consistency test was carried out basing on the results of CNV-seq and MLPA of all 156 cases. Results: Comparing to MLPA, CNV-seq had a coincidence rate of 92.3% (144/156) for DMD gene deletion or duplication, with a sensitivity and positive predictive value of 88.2%, with a specificity and negative predictive value of 94.3%, a missed detection rate of 3.8%, and a Kappa value of 0.839. CNV-seq missed 4 cases with deletions and 2 with duplications due to involved fragments less than 100 Kb, among 20 cases of deletions and 6 cases of duplications detected by MLPA in Group 1. In Group 2, the deletions and duplications detected by CNV-seq were 42% (13/31) and 58% (18/31), respectively, in which the percentage of duplication was higher than that in Group 1. Among those 18 cases with duplications, 3 cases with duplication locating in exon 42~67 were likely pathogenic; while 9 cases with duplication covering the 5' or 3' end of the DMD gene, containing exon 1 or 79 and with only one breakpoint within the gene, along with the last 6 cases with duplications locating at chrX: 32650635_32910000 detected only by CNV-seq, which might be judged as variants of uncertain significance. Conclusions: CNV-seq has a good efficiency to detect fetal DMD gene deletion or duplication in prenatal diagnosis, while a further verification test by MLPA is recommended. The duplications on chrX: 32650635_32910000, 5' or 3' end of DMD gene detected by CNV-seq should be carefully verified and assessed because those variants appear to be nonpathogenic polymorphisms.

The extracellular vesicular pseudogene LGMNP1 induces M2-like macrophage polarization by upregulating LGMN and serves as a novel promising predictive biomarker for ovarian endometriosis recurrence.

STUDY QUESTION: How does ectopic endometrial stromal cell (Ecto-ESC)-derived extracellular vesicular Legumain pseudogene 1 (EV-LGMNP1), a newly identified pseudogene of Legumain (LGMN), contribute to M2-phenotype macrophage polarization, and does it predict recurrence in patients with ovarian endometriosis (EMs)? SUMMARY ANSWER: EV-LGMNP1, which is abundant in Ecto-ESCs and serum from ovarian EMs, can direct macrophages towards an M2 phenotype by upregulating LGMN expression and is a promising biomarker for predicting ovarian EMs recurrence. WHAT IS KNOWN ALREADY: Extracellular vesicles (EVs) can mediate cell-to-cell crosstalk to promote disease progression via cargo molecule transport. Recently, LGMNP1, a newly identified pseudogene of LGMN, has been reported to promote cancer progression by upregulating LGMN. LGMN is a well-studied protein that can induce M2-like polarization. STUDY DESIGN, SIZE, DURATION: An in vitro study was conducted with Ecto-ESCs isolated from ectopic endometrial samples, collected from two patients with ovarian EMs (diagnosed by laparoscopy and histological analysis). A clinical retrospective cohort study of 52 ovarian EMs patients and 21 controls with available preoperative serum samples was carried out (2013-2017). The follow-up period ended either at the time of recurrence or on 31 December 2018. PARTICIPANTS/MATERIALS, SETTING, METHODS: Ecto-ESC-derived EVs (EV/Ecto-ESCs) were characterized by nanoparticle tracking analysis, transmission electron microscopy and western blotting. EV internalization by THP-1 cells, which are the most widely used primary human macrophages model, was detected by fluorescence labelling. After EV treatment, THP-1 cell polarization was detected by quantitative real-time PCR (qRT-PCR) and western blot analyses of CD86 (M1-related marker) and CD206 (M2-related marker). LGMNP1 mRNA expression level in EVs from both primary ectopic endometrioc stromal cells and serum was examined using qRT-PCR. Additionally, the expression of LGMN, the downstream target gene of LGMNP1, in THP-1 cells was evaluated using qRT-PCR and western blotting. Kaplan-Meier and multivariate Cox regression analyses were applied to evaluate the independent predictive factors of EMs recurrence-free survival. A novel nomogram model based on serum EV-LGMNP1 was then formulated to predict EMs recurrence. MAIN RESULTS AND THE ROLE OF CHANCE: In vitro assays demonstrated that EV/Ecto-ESCs drove macrophages towards an M2-like phenotype. Moreover, LGMNP1 contributed to EV/Ecto-ESC-induced M2 macrophage polarization by upregulating LGMN mRNA expression levels. Clinically, serum EV-LGMNP1 was more highly expressed in recurrent EMs patients than in controls and EMs patients without recurrence. Survival analysis and our novel nomogram reconfirmed that serum EV-LGMNP1 was a novel promising and meaningful non-invasive biomarker for predicting EMs recurrence. LARGE SCALE DATA: N/A. LIMITATIONS, REASONS FOR CAUTION: In vitro experiments were only performed on samples from two patients with ovarian endometriosis, and a larger sample size is needed. ESCs isolated from the eutopic endometrium of EMs and non-EMs patients should be studied in the future. Additionally, in vitro experiments should be performed using endometrial epithelium cells and further in vivo experiments, such as using mice endometriotic models to investigate whether EV/Ecto could induce M2 macrophage polarization, should be conducted. Moreover, multicentre, large-sample data are needed to validate our predictive nomogram model. WIDER IMPLICATIONS OF THE FINDINGS: Our study provides novel insights into the mechanism of M2 polarization involved in ovarian EMs progression mediated by an 'EV-shuttled pseudogene LGMNP1' mode. In addition, serum EV-LGMNP1 may serve as a novel non-invasive biomarker for predicting recurrence, providing a new therapeutic target for ovarian EMs. STUDY FUNDING/COMPETING INTEREST(S): This project was supported by funding from the National Natural Science Foundation of China (81971361), the Natural Science Foundation of Shanghai Science and Technology (19ZR1406900), the Shanghai 'Rising Stars of Medical Talent' Youth Development Program (AB83030002019004), the Clinical Research Plan of SHDC (SHDC2020CR4087), the Shanghai Municipal Health Commission (202040498), the Research and Innovation Project of the Shanghai Municipal Education Commission (2019-01-07-00-07-E00050) and the Clinical Research Plan of SHDC (SHDC2020CR1045B). There are no competing interests to declare.

[Preliminary study of the fourth-generation CAR-T cells targeting CS1 in the treatment of refractory and recurrent multiple myeloma].

Objective: To design the fourth-generation chimeric antigen receptor-T (CAR-T) cells that secrete interleukin-7 (IL7) and chemokine C legend 19 (CCL19) on the basis of the second-generation CAR, and to analyze and compare the differences in proliferation, chemotaxis, tumor cell clearance and persistence in the microenvironment of multiple myeloma (MM) between them. Methods: The fourth-generation CAR vector plasmid was constructed by using 2A self-cleaving peptide technology. The third-generation lentiviral packaging system was used to prepare high-titer lentivirus. Flow cytometry was used to monitor the transduction efficiency of lentivirus and the subtype changes of CAR-T cells. The enzyme-linked immunosorbent assay (ELISA) was used to quantify the IL7 and CCL19 secreted by CAR-T cells.The calculation of absolute number of CAR-T cells during culture was used to analysis cell proliferation activity. Transwell migration assay was used to verify the chemotactic ability of CAR-T cells. The specific killing activity of CAR-T cells was detected by using the luciferase bioluminescence method. The NOD-Prkdcem26Cd52Il2rgem26Cd22/Nju (NOD) mouse xenograft model was used to verify the anti-myeloma activity and safety of CAR-T cells in vivo. Results: Flow cytometry results showed that the stable CAR expression rates of the second-generation anti-CS1 CAR-T and fourth-generation anti-CS1-IL7-CCL19 CAR-T cells were (91.50±0.29)% and (46.7±0.12)%, respectively. CAR-T cells were successfully constructed. Subtype analysis demonstrated that the ratio of stem memory T cell (TSCM) in anti-CS1-IL7-CCL19 CAR-T cells was (67.58±0.59)%, which was significantly higher than (50.74 ± 1.01)% of anti-CS1 CAR-T (P=0.000 1), with more strong immune memory function and better durability. Anti-CS1-IL7-CCL19 CAR-T cells can continuously secrete IL7 and CCL19 compared to MOCK-T and anti-CS1 CAR-T (P<0.000 1). The number of anti-CS1-IL7-CCL19 CAR-T cells reached (22.77±0.79)×10(6) on the 9th day after lentivirus transduction, which was significantly higher than (9.40±0.79)×10(6) of anti-CS1 CAR-T cells (P=0.000 1), with stronger proliferation ability. The number of chemotaxis cells of anti-CS1-IL7-CCL19 CAR-T cells to reactive T cells was (109.0±4.04), which was significantly higher than (9.33±1.20) of MOCK-T (P<0.000 1) and (7.33±0.88) of anti-CS1 CAR-T (P<0.000 1), with stronger chemotactic ability. The specific killing activity showed that both anti-CS1-IL7-CCL19 CAR-T and anti-CS1 CAR-T cells had specific killing efficacies when compared with the MOCK-T cells (P<0.000 1). Animal experiment indicated that anti-CS1-IL7-CCL19 CAR-T cells significantly reduced the tumor burden (P<0.000 1) and extended the overall survival time (P=0.006 1) of tumor-bearing mice. Conclusions: The anti-CS1-IL7-CCL19 CAR-T cells designed in this study show stronger proliferative activity, chemotactic ability, and durability without affecting the anti-myeloma activity in vivo and in vivo, which provides strategies for overcoming the defects of low survival rate, poor durability and inhibition by tumor microenvironment of traditional CAR-T cells, and offers preliminary experimental basis for the clinical application of the fourth-generation CAR-T cells.

[Feasibility study of telemedicine model for diagnosis and treatment of patients with obstructive sleep apnea hypopnea syndrome in China].

Objective: To explore the feasibility of applying telemedicine model in disease management for patients with obstructive sleep apnea hypopnea syndrome (OSAHS) in China. Methods: A total of 24 patients were enrolled with suspected OSAHS who were admitted to the Sleep Center of Peking University People's Hospital from October 2015 to September 2016. Patients were diagnosed by electronic questionnaire assessment and home sleep apnea monitoring (HSAT) and were treated with remote automatic positive airway pressure (APAP). After 1 week, 1 month and 3 months of treatment, the patients were followed up by video. The follow-up questionnaire was completed by the patients through an independent data management platform. The APAP treatment data and compliance data were obtained through a built-in digital card of the APAP device. Linear regression model was used to explore the factors related to patient compliance. One-way repeated-measure analysis of variance was used to compare the changes of APAP duration and apnea hypopnea index (AHI) among patients at different treatment time points. Paired t-test was used to compare the EPWORTH scale (ESS) scores before and after treatment. Results: A total of 22 patients were diagnosed with OSAHS, including 20 males (90.9%), aged (45.6±10.2) years and AHI before treatment was (46.9±20.4) times/h. A total of 20 OSAHS patients received APAP treatment, and the proportion of patients with good compliance after 1 week, 1 month and 3 months of treatment were 15/19, 10/19 and 8/18, respectively. The severity of sleepiness before treatment affected compliance. Each 1-point increase in ESS score was associated with a 6.16% (95%CI: 3.01%, 9.31%) increase in compliance. Age, body mass index and AHI before treatment had no effect on compliance (all P values>0.05). The AHI of the patients who had been treated for 1 week, 1 month and 3 months were (2.5±2.1), (2.2±1.6) and (1.9±1.0) times/h, respectively. (P=0.195). After 3 months of treatment, the ESS score was (7.0±3.3), lower than that before treatment (10.6±3.1) (P=0.079). Conclusion: Telemedicine mode of diagnosis and treatment of OSAHS patients has good therapeutic effect and patient compliance, which is practical and feasible.

[Diagnostic value of platelet associated biomarkers in chronic periprosthetic joint infection].

Objective: To evaluate the diagnostic value of platelet count(PC),PC to mean platelet volume(MPV) ratio(PC/MPV) and plateletcrit(PCT) in chronic periprosthetic joint infection(PJI). Method: The medical records of 159 patients who underwent hip or knee revisions at Department of Joint Surgery,Affiliated Hospital of Qingdao University from August 2013 to June 2019 were retrospectively reviewed. There were 51 patients(26 knees and 25 hips) in the PJI group,which included 28 males and 23 females,aged (68.0±11.8)years (range:32 to 84 years)with a body mass index(BMI)of (26.1±3.6) kg/m².There were 116 patients(19 knees and 97 hips) in the aseptic loosening(AL) group,including 67 males and 49 females,aged (70.3±8.9)years(range:49 to 89 years)with a BMI of (25.0±3.6)kg/m².The plasma C-reactive protein(CRP),erythrocyte sedimentation rate(ESR),PC,MPV,PC/MPV and PCT levels of the two groups were recorded and analyzed. Receiver operating characteristic curve was used to calculate the sensitivity and specificity of each biomarker,expect for MPV,and the diagnostic value of each biomarker was compared according to the area under the curve(AUC).Independent-sample t test or Mann-Whitney U test were used for comparison between groups. Result: Compared with AL group,AJI group had significantly higher levels of CRP,ESR,PC,PC/MPV and PCT(all P<0.05),but lower level of MPV (P<0.05).The AUCs for CRP,ESR,PC,PC/MPV and PCT were 0.820, 0.829, 0.689, 0.668 and 0.676,respectively. Based on the Youden index,the optimal predictive cutoff for CRP was 11.12 mg/L,with a sensitivity of 74.4% and a specificity of 87.1%.The optimal predictive cutoff for ESR was 17.60 mm/1 h,with a sensitivity of 81.4% and a specificity of 75.3%.The optimal predictive cutoff for PC was 243.00×109/L,with a sensitivity of 60.6% and a specificity of 71.8%.The optimal predictive cutoff for PC/MPV was 24.95,the sensitivity was 58.1% and the specificity was 74.1%.And the optimal predictive cutoff for PCT was 0.24%,with a sensitivity of 69.8% and a specificity of 63.5%. Conclusion: PC,PC to MPV ratio and PCT were of limited value to diagnose PJI.

[Prognosis analysis of local recurrence after excision of breast phyllodes tumors].

Objective: To examine treatment outcomes of breast phyllodes tumors and the prognosis factors of local recurrence. Methods: This retrospective cohort study included 276 patients who underwent surgical resection at Breast Center, Peking University People's Hospital from January 2011 to December 2019. Tumor subtype and histopathological features were determined from pathology reports, and the deadline of follow-up was September 30th, 2020. All 276 patients underwent open surgery, including 17 patients of mastectomy, and 259 patients of lumpectomy. The enrolled patients were all female, with age of (41.5±11.3) years (rang: 11 to 76 years), and tumor diameter of 35(28) mm (M(QR)). The Kaplan-Meier method and Log-rank test were used for survival analysis. The multivariate analysis was implemented using the Cox proportional hazard model. Results: According the pathologic test, there were 191 patients of benign phyllodes tumor, 67 patients of borderline tumor and 18 patients of malignant tumor. There were 249 patients with a follow-up of more than 6 months, and 14.1% (35/249) had local recurrence. The time-to-recurrence was (28.6±22.2) months (range: 2 to 96 months), (29.1±18.1) months (range: 2 to 80 months), (32.1±30.1) months (range: 5 to 96 months) and (12.0±6.9) months (range: 8 to 20 months) for benign, borderline and malignant phyllodes tumors. Tumor diameter (≥100 mm vs.<50 mm, HR=3.968, 95%CI: 1.550 to 10.158, P=0.004) and malignant heterologous element (yes vs. no, HR=26.933, 95%CI: 3.105 to 233.600, P=0.003) were prognosis factors of local recurrence. One death from malignant phyllodes occurred after distant metastasis. The 3-year disease-free survival rates of benign, borderline and malignant phyllodes tumor were 88.2%, 81.7% and 81.4% (P=0.300). Conclusion: Phyllodes tumors have a considerable local recurrence rate, which may be associated with tumor diameter and malignant heterologous element.

[Expression of CYP2E1 in different brain regions during the formation of toxic cerebral edema induced by 1, 2-DCE in mice and its effect on cerebral edema formation].

Objective: To study the expression changes of cytochrome P-450 2E1 (CYP2E1) in different brain regions during 1, 2-dichloroethane (1, 2-DCE) exposure, and to explore the effect of CYP2E1 on the influence and effect of brain edema caused by 1, 2-DCE poisoning. Methods: In December 2018, forty female SPF Kunming mice were randomly divided into 4 groups: control group, 1 d, 2 d, and 3 d exposure groups, with 10 mice in each group. Placed in a 100 L static poisoning cabinet (5 animals/cabinet) , inhaled and exposed to the poison at 1.2 mg/L 1, 2-DCE for 3.5 h per day for 1 d, 2 d and 3 d. Except for exposure to 1, 2-DCE, other treatment methods were the same as those in the exposure groups. They were put to death the next day after the end of the poisoning, and the brain tissue was quickly removed and divided into sections. HE staining method was used for different brain regions. Western blotting method was used to detect the protein content of CYP2E1, occludin and claudin5 in different brain regions, real-time fluorescent quantitative PCR method was used to detect the mRNA expression levels of CYP2E1, occludin and claudin5. Histopathological observations were performed, and the kit method was used to detect malondialdehyde (MDA) , reduced glutathione (GSH) content and catalase (CAT) activity in different brain regions. The experimental data were analyzed by SPSS 20.0. One-way analysis of variance (one-way ANOVA) was used for the comparison of multiple groups, and the SNK (q test) method was used for the pairwise comparison between groups.P<0.05 was considered statistically significant. Results: Compared with the control group, histopathological observations showed obvious brain edema in the 2 d and 3 d exposure groups; Compared with the control group, the MDA content, CYP2E1 protein and mRNA expression levels in the cerebellar tissues of the mice in the 3 d exposure group were significantly increased, and the differences were statistically significant (P<0.05) ; Compared with the control group, the GSH content, CAT activity, occludin and claudin5 protein expression levels in the cerebellar tissues of the mice in each exposure group were significantly reduced, and the differences were statistically significant (P<0.05) . There was no significant difference in the above indicators of frontal cortex in each group in mice (P>0.05) . Conclusion: 1, 2-DCE can induce the expression of CYP2E1 in cerebellar tissues of mice, and cause oxidative damage and brain edema, but has no effect on the expression of CYP2E1 in frontal cortex of mice.

Research Progress of Aquatic Corpse Decomposition and Postmortem Submersion Interval Estimation.

ABSTRACT: Human corpses can be found in a variety of aquatic environments. The decomposition of corpses in aquatic environments is different from those on land. A large number of factors influence the decomposition process in water, therefore postmortem submersion interval （PMSI） is difficult to estimate. To date, while studies on aquatic corpses are obviously fewer than those on terrestrial corpses, there are many problems in practical work. This review summarizes the stages and influencing factors of aquatic corpse decomposition, and introduces the relevant research progress of PMSI estimation based on decomposition stages, postmortem phenomena, aquatic insects, biofilm, and physical and chemical methods, in order to provide reference for aquatic decomposition researches and practices.

Prospects of Application of Microbial Biofilm to Estimate PMI of Corpses in Water.

ABSTRACT: In forensic pathology, the estimation of postmortem interval （PMI） has always been a difficult issue, and there is still lack of effective methods to estimate PMI of corpses in water. Microbial biofilm refers to the microbial population attached to non-biological or biological surfaces by microorganisms during microbial growth, that has a three-dimensional structure, surrounded by extracellular polymers and matrix networks created by itself. A series of community succession phenomena of microorganisms occur during the occurrence and development of microbial population. The microbial community and its succession process of this kind of biofilm attached to the surface of a corpse in water may become a new basis for estimation of the PMI of corpses in water. This review elucidates on the concept, classification, research methods, and influencing factors of biofilm and analyzes its application prospects in PMI estimation of corpses in water, which would provide new ideas for the researches in this field.

Epigallocatechin gallate enhances treatment efficacy of oral nifedipine against pregnancy-induced severe pre-eclampsia: A double-blind, randomized and placebo-controlled clinical study.

WHAT IS KNOWN AND OBJECTIVE: Oral nifedipine is commonly used to treat pre-eclampsia, one of the most severe complications during pregnancy, but its clinical efficacy is less than ideal. Epigallocatechin gallate (EGCG), a natural compound from green tea, could benefit cardiovascular health especially hypertension. We investigated the clinical efficacy of EGCG, when complemented with oral nifedipine, in treating pre-eclampsia. METHODS: A total of 350 pregnant women with severe pre-eclampsia were recruited and randomized to receive oral nifedipine, together with placebo (NIF+placebo) or EGCG (NIF+EGCG). The primary treatment outcome was the time needed to control blood pressure and interval time before a new hypertensive crisis, whereas the secondary treatment outcome was the number of treatment doses to effectively control blood pressure, maternal adverse effects and neonatal complications. RESULTS AND DISCUSSION: Comparing NIF+EGCG group to NIF+placebo group, the time needed to control blood pressure was significantly shorter (NIF+EGCG 31.2±16.7 minutes, NIF+placebo 45.3±21.9 minutes; 95% CI 9.7-18.5 minutes), whereas interval time before a new hypertensive crisis was significantly prolonged (NIF+EGCG 7.2±2.9 hours, NIF+placebo 4.1±3.7 hours; 95% CI 2.3-3.9 hours), and the number of treatment dosages needed to effectively control blood pressure was also lower. Between the two treatment groups, no differences in incidence rates of maternal adverse effects or neonatal complications were observed. WHAT IS NEW AND CONCLUSIONS: EGCG is both safe and effective in enhancing treatment efficacy of oral nifedipine against pregnancy-induced severe pre-eclampsia, but formal validation is required prior to its recommendation for use outside of clinical trials.

[Cryptococcus bloodstream infection: a retrospectively analysis of 16 cases in one tertiary hospital].

Objective: To explore the clinical characteristics of cryptococcus bloodstream infection. Methods: A retrospectively analysis was performed by collecting data of clinical manifestations, underlying diseases, susceptible factors, therapy and prognosis of cryptococcus bloodstream infection in the First Affiliated Hospital of Soochow University from December 2006 to December 2017. Results: The most common symptom of the 16 patients was fever (12/16). When combined with cryptococcus infection of central nervous system, the manifestations may include dizziness, headache, nausea and vomiting.Among the 16 patients, 11 cases had underlying diseases, which listed as leukemia, lymphoma, diabetes mellitus, hepatitis B, cirrhosis, chronic kidney disease and carcinoma of the liver; 12 cases had invasive procedures, 6 cases had hypoproteinemia, 4 cases had long-term usage of glucocorticoids, and 5 cases received broad spectrum antibiotics longer than 1 week.8 cases died, 6 cases improved and 2 cases were transferred to the specialized hospital for their newly diagnosed AIDS. Conclusions: Bloodstream infection of cryptococcus is a relatively rare disease.Fever is one of the most common symptoms of cryptococcus bloodstream infection.Underlying diseases, invasive procedures, hypoproteinemia, long-term application of glucocorticoids and antibiotics are the risk factors for cryptococcus bloodstream infection.

[Safety and efficacy of rotational atherectomy in the interventional treatment of coronary chronic total occlusion lesions].

Objective: To investigate the safety and efficacy of rotational atherectomy in the interventional treatment of coronary chronic total occlusion lesions. Methods: In this retrospective study,a total of 31 consecutive patients with coronary chronic total occlusion(CTO) lesions underwent rotational atherectomy in our hospital from February 2004 to December 2016 were enrolled,and the clinical features were analyzed. Coronary atherectomy was performed if balloon failed to cross the CTO lesions or balloon could not be fully dilated in the CTO lesions after wire crossing. The definition of procedure success was defined as residual stenosis less than 20% after implantation of drug eluting stent and rotational atherectomy. After the procedure, the patients were followed up to observe major adverse cardiac and cerebral vascular events which including cardiogenic death, myocardial infarction, cerebrovascular accident, and target lesion revascularization. Results: The 1.25 mm diameter burr was firstly selected in 80.6% (25/31) patients,and 96.8%(30/31) patients used only 1 burr to complete the rotational atherectomy procedure. The complication rate was 9.8% (3/31) including 1 patient with coronary dissection and 3 patients with slow flow or no flow. There was 1 patent with both coronary dissection and slow flow. The procedure success rate was 96.8%(30/31). Interventional treatment related myocardial infarction occurred in 3 patients during hospitalization.The 30 patients with procedure success were followed up 36(11, 96) months. The incidence rate of major adverse cardiac and cerebral vascular events was 13.3% (4/30), of which the cardiogenic death rate was 3.3% (1/30), the myocardial infarction rate was 6.7% (2/30), cerebrovascular accident rate was 3.3%(1/30),and the target lesion revascularization rate was 6.7% (2/30). Conclusion: Rotational atherectomy is safe and effective in the interventional treatment of coronary CTO lesions.

Application of High-throughput Sequencing in Researches of Cadaveric Microorganisms and Postmortem Interval Estimation.

Necrobiome is the main factor causing the cadaver decomposition. Studying the microbial succession during decomposition is one of the main tasks of forensic microbiology. The interactive relationships among cadaver, environment and microorganisms are complicated. The microbial succession study relies on macroscopic monitoring of community composition and the diversity change in each decomposition stage. With the maturity and development of high-throughput sequencing （HTS）, the structure and diversity of microbial communities in different environments have been successively revealed. A new breakthrough to explore the cadaveric microorganisms has been opened as well. It has become the research hotspots in forensic microbiology to reveal the microbial succession in the process of cadaver decomposition and to interpret the essence of various decomposition phenomena by using HTS, which can provide a new reference for postmortem interval （PMI） estimation. The present paper reviews studies on PMI estimation by using cadaveric microorganism. Problems and application prospects of forensic microbiology studies are discussed on the basis of the current application of HTS technology in the exploration of microbial succession.

Au20Si12: A hollow Catalan pentakis dodecahedron.

A stable hollow Au20Si12 cage with Ih symmetry has been predicted using first-principles density functional theory. The stability of the cage-like Au20Si12 structure is verified by vibrational frequency analysis and molecular dynamics simulations. A relatively large highest occupied molecular orbital-lowest unoccupied molecular orbital gap of 1.057 eV is found. Electronic structure analysis shows that clearly p-d hybridizations between Si atoms and Au atoms are of great importance for the stability of Au20Si12 cage. The cage-like Au20Si12 structure may have potential applications in semiconductor industry and microelectronics.

[Analysis on the novel compound heterozygous mutation Fâ ª of a patient with hereditary factor â ª deficiency].

Objective: To investigate the clinical phenotype and genotype characteristics of a Chinese hereditary factor Ⅺ deficiency pedigree. Methods: The activated partial thromboplastin time (APTT), prothrombin time (PT), FⅪ activity (FⅪ: C) were measured by clotting method using automatic coagulation analyzer. The FⅪ antigen (FⅪ: Ag) was assayed by enzyme-linked immunosorbent assay (ELISA). Fifteen exons of F11 from the proband and his pedigree members were amplified by polymerase chain reaction (PCR), then sequenced. Pymol software was used to analyze the novel mutations. Results: APTT, FⅪ: C and FⅪ: Ag of proband was 74.2 s, 4.0% and 2.9%, respectively. For his older sister, APTT, FⅪ: C and FⅪ: Ag was 67.1 s, 3.0% and 1.8%, respectively. APTT, FⅪ: C and FⅪ: Ag of healthy controls were 34.5 s, 100.0% and 100.0%. FⅪ: C of proband's father, mother and brother were 72.0%, 62.0%, and 78.0%, respectively. FⅪ: Ag of them were 50.0%, 43.0%, and 51.8%, respectively. The other coagulant parameters of the proband and his pedigree were all in the normal range. Sequence analysis showed two heterozygous gene mutations in F11 of the proband and his older sister. One was a deletion of T at nucleotide 1 491 in exon 12, resulting in a frameshift. A substitution of leucine 465 by tryptophan and a terminal coden after 7 amino acid: F11NM_13142c.1491delT (p.Leu465Trp.fs*7). The other was a G to A substitution at nucleotide 1 815 in exon 15, resulting in a substitution of glycine 573 by aspartic acid: F11 NM_13142c.1815G>A (p.Gly573Asp). F11NM_13142c.1491delT (p.Leu465Trp.fs*7) heterozygotes were found both in the proband's father and his brother while p. Gly573Asp heterozygote was only found in his mother. F11 of the proband's uncle was wild. Conclusion: The novel compound heterozygous mutations of F11NM_13142c.1491delT (p.Leu465Trp.fs*7) and F11 NM_13142 c. 1815G>A (p.Gly573Asp) are responsible for FⅪ deficiency to the proband, which induced the decrease of FⅪ: C and FⅪ: Ag.

[Association of HLA-DQ and IFNL4 polymorphisms with hepatitis B virus infection and clearance].

Objective: To confirmed the polymorphisms of HLA-DQ and IFNL4 were associated with HBV infection and clearance in a Chinese population. Methods: The Sequenom MassARRAY MALDI-TOF system was used to genotype the HLA-DQrs9275319 and IFNL4rs368234815, rs12971396, rs12979860, and rs8099917. A binary logistic regression test was conducted to estimate the relative risk of these SNPs with HBV infection and clearance. Haploview4.2 software and PHASE software (v2.0.2) were employed to analyze linkage disequilibrium (LD) and haplotype frequencies. The MDR program was applied to analyze interactions between SNP and SNP.Statistical analysis was performed using SPSS 19.0 and P-values were corrected by Bonferroni's corrections. Results: A total of 1,069 subjects were recruited and divided into three groups: 238 healthy controls(HC), 397 with HBV-related chronic liver disease (CLD), 434 with spontaneous clearance (SC). The rs9275319TT was most frequently identified among all groups(86.2% in the CLD group, 77.6% in the SC group, and 75.9% in the HC group).Carriage of the rs9275319 C allele was a protective factor for chronic HBV infection (the allele model: P = 0.000 3, OR,0.514; 95% CI, 0.359-0.738) and clearance (the allele model: P = 0.002, OR, 1.659; 95% CI, 1.197-2.300). HLA-DQ rs9275319 showed a significant association with HBV infection (allele model, OR, 0.514; 95% CI, 0.359-0.738, adjusted P = 0.000 3) and spontaneous clearance (allele model, OR, 1.659; 95% CI, 1.197-2.300, adjusted P = 0.002). However, there was no association between IFNL4 polymorphism and HBV infection((allele model: P = 0.082 for rs368234815; P = 0.063 for rs12971396; P = 0.517 for rs12979860; P =0.695 for rs8099917) or spontaneous clearance ((allele model: P = 0.358 for rs368234815; P = 0.105 for rs12971396; P = 0.640 for rs12979860; P = 0.640 for rs8099917;all P > 0.05). The multifactor dimensionality reduction (MDR) test showed there was a three-way interaction (rs12971396, rs12979860, and rs9275319) between IFNL4 and HLA-DQ polymorphisms for HBV infection (permutation P = 0.009 for the best factor model) and clearance (permutation P = 0.014 for the best factor model). Conclusion: The SNP-SNP interaction between HLA-DQ and IFNL4 is associated with the regulation of HBV infection and natural clearance.

[Study the impacts of diagnosis on occupational noise-induced deafness after bring into the different high frequency hearing threshold weighted value].

Objective: Study of the results and the degree on occupational noise-induced deafness in-to the different high frequency hearing threshold weighted value, in order to provide theoretical basis for the re-vision of diagnostic criteria on occupational noise-induced deafness. Methods: A retrospective study was con-ducted to investigate the cases on the diagnosis of occupational noise-induced deafness in Guangdong province hospital for occupational disease prevention and treatment from January 2016 to January 2017. Based on the re-sults of the 3 hearing test for each test interval greater than 3 days in the hospital, the best threshold of each frequency was obtained, and based on the diagnostic criteria of occupational noise deafness in 2007 edition, Chi square test, t test and variance analysis were used to measure SPSS21.0 data, their differences are tested among the means of speech frequency and the high frequency weighted value into different age group, noise ex-posure group, and diagnostic classification between different dimensions. Results: 1. There were totally 168 cases in accordance with the study plan, male 154 cases, female 14 cases, the average age was 41.18 ±6.07 years old. 2. The diagnosis rate was increased into the weighted value of different high frequency than the mean value of pure speech frequency, the weighted 4 kHz frequency increased by 13.69% (χ(2)=9.880, P=0.002) , 6 kHz increased by 15.47% (χ(2)=9.985, P=0.002) and 4 kHz+6 kHz increased by15.47% (χ(2)=9.985, P=0.002) , the difference was statistically significant. The diagnostic rate of different high threshold had no obvious differ-ence between the genders. 3. The age groups were divided into less than or equal to 40years old group (A group) and 40-50 years old group (group B) , there were higher the diagnostic rate between high frequency weighted 4 kHz (A group χ(2)=3.380, P=0.050; B group χ(2)=4.054, P=0.032) , weighted 6 kHz (A group χ(2)=6.362, P=0.012; B group χ(2)=4.054, P=0.032) , high frequency weighted 4 kHz+6 kHz (A group χ(2)=6.362, P=0.012; B group χ(2)=4.054, P=0.032) than those of speech frequency average value in the same group on oc-cupational noise-induced deafness diagnosis rate, the difference was statistically significant. There was no sig-nificant difference between age groups (χ(2)=2.265, P=0.944) . 4. The better ear's mean value of pure speech fre-quency and the weighted values into different high frequency of working years on each group were compared, working years more than 10 years group was significantly higher than that of average thresholds of each frequen-cy band in 3-5 group (F=2.271, P=0.001) , 6-10 group (F=1.563, P=0.046) , the difference was statistically significant. The different high frequency weighted values were higher than those of the mean value of pure speech frequency, and the high frequency weighted 4 kHz+6 kHz had the highest frequency difference, with an average increase of 2.83 dB. 5. The diagnostic rate into weighted different high frequency was higher in the mild, moderate and severe grades than in the pure speech frequency. In the comparison of diagnosis for mild occupational noise-induced deafness, in addition to the weighted 3 kHz high frequency (χ(2)=3.117, P=0.077) had no significant difference, the weighted 4 kHz (χ(2)=10.835, P=0.001) , 6 kHz (χ(2)=9.985, P=0.002) , 3 kHz+4 kHz (χ(2)=6.315, P=0.012) , 3 kHz+6 kHz (χ(2)=6.315, P=0.012) , 4 kHz+6 kHz (χ(2)=9.985, P=0.002) , 3 kHz+4 kHz+6 kHz (χ(2)=7.667, P=0.002) were significantly higher than the diagnosis rate of the mean value of pure speech frequency. There was no significant difference between the two groups in the moderate and se-vere grades (P>0.05) . Conclusion: Bring into different high frequency hearing threshold weighted value in-creases the diagnostic rate of occupational noise-induced deafness, the weighted 4 kHz, 6 kHz and 4 kHz+ 6 kHz high frequency value affects the result greatly, and the weighted 4 kHz+6 kHz high frequency hearing threshold value is maximum the effect on occupational noise-induced deafness diagnosis.

[Effect of mouse nerve growth factor on cognitive impairment in whole brain irradiation rats].

OBJECTIVE: To investigate the positive effects of mouse nerve growth factor(mNGF) on cognitive impairment in whole brain irradiation rats. METHODS: Fifty-five male Sprague-Dawley rats were randomly divided into normal control group, mNGF treated group, saline treated group.mNGF treated group and saline treated group were cranial irradiated at a single dose of 12 Gy by X-ray.30 days after radiation each group were treated with correspondent drugs.60 days after radiation, Morris water maze experiment, EB leakage of the brain, and expressions of neuN, vWF, ZO-1 in hippocampus by immunofluorescence, and expressions of neuN, vWF, ZO-1, VEGF and GFAP in hippocampus by Western blot were tested and analyzed. RESULTS: The escape latencies: normal control group< mNGF treated group< saline treated group; the numbers of crossing hidden platform in these 3 groups were 3.00± 1.08, 1.50± 1.08, 0.38± 0.48 times (P<0.01). EB leakage of these 3 groups were 0.14±0.14, 0.66±0.20 and 1.36±0.27 µg/g (P<0.05). In immunofluorescence, expressions of neuN, vWF and ZO-1: normal control group> mNGF treated group> saline treated group.In Western blot, expressions of neuN, vWF and ZO-1: normal control group> mNGF treated group> saline treated group, yet the expressions of VEGF and GFAP: normal control group< mNGF treated group< saline treated group. CONCLUSION: mNGF ameliorates cognitive impairment after whole brain irradiation.

Genetic Polymorphisms of 21 STR Loci in Hunan Province-based Han Population.

OBJECTIVES: To investigate the genetic polymorphisms of 21 short tandem repeat （STR） loci （D3S1358, D13S317, D7S820, D16S539, Penta E, D2S441, TPOX, TH01, D2S1338, CSF1PO, Penta D, D10S1248, D19S433, vWA, D21S11, D18S51, D6S1043, D8S1179, D5S818, D12S391 and FGA）. METHODS: A total of 560 blood samples were collected from unrelated healthy individuals of Han population in Hunan Province. Chelex-100 extraction method was applied to the extraction of genomic DNA, and an AGCU EX22 Kit and 9700 STR amplification was used in amplification reactions. The products were separated and analyzed on 310 Genetic Analyzer. RESULTS: A total of 248 alleles were observed, the allelic frequencies ranging from 0.001 to 0.518. Observation of genotype distributions for each locus showed no deviations from Hardy-Weinberg equilibrium except Penta E （P=0.023）. The combined power of discrimination, combined power of exclusion, and combined matching probability of the 21 STR loci were approximately 0.999 999 999 999 999 999 999 999 8, 0.999 999 998, and 1.36×10⁻²5, respectively. CONCLUSIONS: The 21 STR loci show high polymorphisms in the Han population, which can provide valuable data and a theoretical basis for forensic individual identification and paternity testing.